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Study of the One-Step Growth Curve of Equine Infectious Anemia Virus by Immunofluorescence

^a Department of Veterinary Pathology, College of Veterinary Medicine, Washington State University, and the Veterinary Science Research Division, Agricultural Research Service, U.S. Department of Agriculture, Pullman, Washington 99163

ABSTRACT

Primary horse leukocyte cultures were inoculated with 2 or 10 50% tissue culture infective doses (TCID₅₀) of equine infectious anemia (EIA) virus per cell, and the titer of cell-associated and fluid-phase virus was determined from 1 to 72 hr postinoculation (PI). Cover slips were collected from 4 to 72 hr PI and stained for EIA viral antigen by the indirect immunofluorescent (FA) technique. Viral replication was detected after a latent period of approximately 18 to 24 hr and reached peak titers of approximately 10^4.5 to 10⁶ TCID₅₀/0.5 ml from 48 to 72 hr PI. The fluid phase contained 10¹ to 10² TCID₅₀/0.5 ml more virus than the cells. Viral antigen was first detected by FA from 18 to 24 hr PI. Approximately 75% of the cells contained antigen in their cytoplasm 72 hr PI. The FA technique is a sensitive method for detecting EIA virus in horse leukocyte cultures.

¹ Present address: National Institute of Animal Health, Kodaira, Tokyo, Japan.

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